Professional Documents
Culture Documents
Rohit sah
PRESENTATION TITLE Introduction
• Liver function tests (also known as a liver panel) are blood tests that measure
different enzymes, proteins, and other substances made by the liver.
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PRESENTATION TITLE Classification
I. Tests based on abnormalities of bile pigment metabolism:
• Serum bilirubin and VD Bergh reaction
• Urine bilirubin
• Urine and fecal urobilinogen
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PRESENTATION TITLE
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PRESENTATION TITLE Serum bilirubin and VD Bergh reaction
Principle:
• Methods for detecting and estimating bilirubin in serum are based on the formation of a
purple compound azo-bilirubin where bilirubin in serum is allowed to react with a
freshly prepared, solution of VD Bergh’s diazo-reagent, This reaction is called VD
Bergh Reaction.
• VD Bergh’s diazo-reagent;
It consists of two solutions:
Solution A: Dissolve 1 g sulphanilic acid in 15 ml conc. HCl; and make volume 1L with
distilled water.
Solution B: 0.5% sodium nitrite in water.
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Mix 5 ml solution A in 0.15 ml solution B (prepare fresh).
PRESENTATION TITLE
Procedure
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PRESENTATION TITLE
PRESENTATION TITLE
Clinical Significance
• In haemolytic (prehepatic) jaundice,
there is a moderate increase in serum total and indirect bilirubin concentration.
It may occur due to haemoglobinopathies or Rh incompatibility in children,
malaria or due to toxic drug reactions.
• Obstructive jaundice
There is markedly increased in indirect bilirubin.
It may occur due to carcinoma on the head of the pancreas, gallstones or other
causes. In obstructive jaundice indirect bilirubin level markedly increased.
• In hepatic jaundice,
There is increased in direct bilirubin.
It may be due to defective conjugation as in chronic hepatitis, Gilbert’s disease
11 and Criggler-Najjar’s syndrome.
PRESENTATION TITLE Urinary Bilirubin
• Normally bilirubin is absent in urine.
• This can be detected by Fouchet's test
Fouchet's test
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PRESENTATION TITLE
Interpretations
• Unconjugated bilirubin is insoluble in water and always absent in urine
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PRESENTATION TITLE Urinary urobilinogen
• Normally trace amounts is present in urine.
• Average is 0.64 mg, maximum normal 4 mg/ 24 hours.
• This can be detected by Erhlich’s test.
Principle:
• Urobilinogen + p-dimethylamino benzaldehyde (Erhlich’s reagent) = Form red
color.
• Ascorbic acid is added as a reducing agent to keep urobilinogen in a reduced
state.
Procedure
• The addition of saturated sodium acetate stops the reaction.
• Take two test tubes.
• Label one as a test (T) and the other for comparison (B).
• Add 5 ml of freshly voided urine to each test tube.
• Add 0.5 mL of Ehrilich reagent to test (T).
• Add 0.5 mL of conc HCL to the B test tube and mix.
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• Leave both test tubes at room temperature for 5 minutes.
• Note for the red color in the test (T) tube.
PRESENTATION TITLE
Interpretations
• In obstructive jaundice no urobilinogen is present in urine because bilirubin cannot
enter intestine.
Interpretations
• In Hemolytic jaundice faecal urobilinogen is increased which is dark colored
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PRESENTATION TITLE Galactose Tolerance Test
• The normal liver is able to convert galactose into glucose but this function is impaired
in intrahepatic diseases and the amount of blood galactose and galactose in urine is
excessive.
Methods:
This can be of two types:
1. Oral galactose tolerance test (Maclagan) and
2. IV galactose tolerance test.
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PRESENTATION TITLE Oral galactose tolerance test (Maclagan)
• The test is performed in the morning after a night’s fast. A fasting blood sample is
collected which serves as “control”. 40 gm of galactose dissolved in a cup-full of
water is given orally.
• Further blood samples are collected at 1/2 hourly intervals for two hours (similar to
GTT).
Interpretations
Interpretations
• A normal response should have a curve beginning on the average at about 200 mg
galactose/100 dl, falling steeply during the one hour and reaching a figure between 0 to
10 mg% by end of 2 hours.
• In most cases of obstructive jaundice similar results are obtained, unless there is
parenchymal damage.
• In parenchymatous diseases with liver cell damage, the fall in blood galactose takes
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place more slowly. Normally no galactose is detected in 2½ hours sample, but in
parenchymatous disease, value is greater than 20 mg/dl.
PRESENTATION TITLE
Fructose Tolerance Test
Interpretations
• Normal response shows little or no rise in the blood sugar level. The highest blood
sugar value reached during the test should not exceed the fasting level by more than
30 mg%.
• Similar result is obtained in most cases of obstructive jaundice cases (provided no
parenchymal damage).
21 • In infectious hepatitis and parenchymatous liver cells damage: Rise in blood sugar
is greater than above, but the increases obtained are never very great.
Epinephrine Tolerance Test (Storage Function)
PRESENTATION TITLE
Principle:
• The response to epinephrine as evidenced by elevation of blood sugar is a manifestation
of glycogenolysis and is directly influenced by glycogen stores of liver.
Method
• The patient is kept on a high carbohydrate diet for three days before the test. After an
overnight fast, the fasting blood sugar is determined.
• 0.01 ml of a 1 in 1000 solution of epinephrine per kg body weight in injected.
• The blood sugar is then determined in samples collected at 15 minutes intervals up to
one hour.
Interpretations
• Normally in the course of an hour, the rise in blood sugar over the fasting level exceeds
by 40 mg% or more.
• In parenchymal hepatic diseases the rise is less.
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• It is of much use for diagnosis of glycogen storage diseases, specially in von Gierke’s
disease, in which blood glucose rise is not seen due to lack of glucose6-phosphatase.
PRESENTATION TITLE
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Determination of Total Plasma Proteins and Albumin and Globulin and
A:G Ratio
PRESENTATION TITLE
• Liver is the site of albumin synthesis and also possibly of some of α-and β-globulins.
• This yields most useful information in chronic liver diseases.
• This can be detected by Biuret test.
Biuret Test
Principle ;
• Proteins, which contain peptide linkages from a complex with copper in alkaline
medium giving a violet colour and this reaction, is called the biuret reaction.
• The intensity of this colour is proportional to the number of peptide linkages present
and thus is a measure of the concentration of proteins.
• Albumins are estimated in serum using the biuret reaction after precipitation and
separation of serum globulins by sodium sulphate. Globulins are precipitated by
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sodium sulphate.
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PRESENTATION TITLE
Calculation
Normal range
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PRESENTATION TITLE
PRESENTATION TITLE
Interpretations
• In obstructive jaundice normal values are the rule, as long as the obstructive
jaundice is not associated with accompanying liver cell damage.
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PRESENTATION TITLE Estimation of Plasma Fibrinogen
• Fibrinogen is formed in the liver and likely to be affected if considerable liver
damage is present.
• Values below 100 mg% have been reported in severe parenchymal liver damage.
• Such a situation is found in severe acute insufficiency such as may occur in:
Acute hepatic necrosis
Poisoning from carbon tetrachloride
In advanced stages of liver of
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PRESENTATION TITLE
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Determination of serum cholesterol and ester cholesterol and their
ratio
PRESENTATION TITLE
• The liver plays an important and active role in the metabolism of cholesterol
including its synthesis, esterification, oxidation and excretion.
Interpretations.
• Normal total blood cholesterol ranges 150 to 250m/dl and abt 60-70% in esterified
form.
• In/obstructive jaundice, total blood cholesterol level is increased and the esterified
form is also increased.
• In liver damage, no change in total blood cholesterol level but its esterified form is
decreased
• In severe acute hepatic necrosis, total blood cholesterol level is decreased upto
100mg/dI and there is marked reduction in % present as esters.
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PRESENTATION TITLE
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PRESENTATION TITLE
Hippuric acid test
Principle
• Best known test for the detoxicating function of liver.
• Liver removes benzoic acid, administered as sodium benzoate, either orally or IV, and
combines with amino acid glycine to form hippuric acid. The amount of hippuric acid
excreted in urine in a fixed time is determined. (Refer chapter on detoxication).
• The test thus depends on two factors (i) The ability of liver cells to produce and
provide sufficient glycine and (ii) The capacity of liver cells to conjugate it with the
benzoic acid.
• For reliable result, renal function must be normal. If there is any reason to suspect
renal impairment, a urea clearance test should be done simultaneously.
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Methods: Both oral and IV forms of the hippuric acid test are in use.
PRESENTATION TITLE Oral hippuric acid test:
Procedure
• Dissolve 6.0 gm of sodium benzoate in approx 200 ml of water.
• The test may be started 3 hours after a light breakfast of toast and tea. Food should
not be given until late in the test.
• The patient is allowed to drink the sodium benzoate solution and time is noted.
• Any urine passed during this 4 hours is kept and added to that passed at the end of 4
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hours. The amount of hippuric acid excreted in this 4 hours period is estimated.
PRESENTATION TITLE
Interpretations
• Smaller amounts are found when there is either acute or chronic liver damage.
Amounts lower than 1.0 gm may be excreted by patients with infectious hepatitis.
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PRESENTATION TITLE
IV Hippuric Acid Test ;
Procedure
• 1.77 gm of sodium benozate dissolved in 20 ml of DW as a sterile solution given
• IV. Shortly before the injection, the patient empties the bladder, which is discarded.
• The bladder is emptied after one hour and two hours after the injection.
Interpretations
• In normal health, hippuric acid equivalent to at least 0.85 gm of sodium benzoate,
or to 0.7 gm of benzoic acid should be excreted in the one hour, or equivalent to
37 1.15 gm of benzoic acid in the first two hours.
• Excretion of smaller amounts more than above indicate the presence of liver
PRESENTATION TITLE BSP Retention Test (Bromsulphthalein
Test)
Principle
1. The ability of the liver to excrete certain dyes, e.g. BSP is utilised in this test.
3. Removal of BSP by the liver involves conjugation of the dye as a mercaptide with
the cysteine component of glutathione. The reaction of conjugation of BSP with
glutathione is rate-limiting, and thus it exerts a controlling influence on the rate of
removal of the dye.
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Procedure
• With the patient fasting, inject IV slowly, an amount of 5 per cent BSP solution, which
PRESENTATION TITLE
• Withdraw 5 to 10 ml of blood, 25 and 45 minutes after the injection and allow the
specimens to clot.
• Separate the sera and estimate the amount of the dye in each sample.
Interpretations
• In normal healthy individual not more than 5 per cent of the dye should remain in the
blood at the end of 45 minutes. The bulk of the dye is removed in 25 minutes and less
than 15 per cent is left at the end of 25 minutes.
• Interpretation: Normally 50 per cent or more of the dye disappears within 8 minutes
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PRESENTATION TITLE Bilirubin tolerance test:
• 1 mg/kg body weight of bilirubin is injected IV. If more than 5 per cent of the injected
bilirubin is retained after 4 hours, the excretory and conjugating function of the liver is
considered abnormal.
• The bilirubin excretion test has been recommended by some authorities as a better test
of excretory function of the liver as compared to dye tests as bilirubin is a normal
physiologic substance and the dyes are foreign to the body.
• But the test is not used routinely and extensively due to its high cost.
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PRESENTATION TITLE
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PRESENTATION TITLE
Principle
• The prothrombin time is the time required for
the plasma to clot after addition of tissue factor
(thromboplastin) and an optimal concentration of
calcium.
• This Indicates overall efficiency of extrinsic
pathway.
Name of the method :
Quick’s method
Specimen :
Citrated Plasma
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PRESENTATION TITLE
Procedure
1. Take 0.1 ml plasma in clean test tube. The plasma should be pre-warmed at
37°C in water bath .
2. Add 0.1 ml tissue thromboplastin, mix well and incubate at 37°C for 2 minutes.
3. Add 0.1 ml calcium chloride solution, mix and start the stop watch.
4. At the first appearance of a fibrin clot, stop the watch immediately.
5. Record the time.
6. Report prothrombin time in seconds.
Reference 12 to 15 seconds
range;
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PRESENTATION TITLE
Interpretations
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PRESENTATION TITLE
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Determination of blood NH3
PRESENTATION TITLE
• Ammonia, also known as NH3, is a waste product made by your body during the
digestion of protein. Normally, ammonia is processed in the liver, where it is
changed into another waste product called urea. Urea is passed through the body
in urine.
Procedure
• The patient should come for the test after overnight 12 hours fast, only small amounts of fluids can be
taken during that time.
• Take fasting specimen of blood for NH3 determination.
• After that, give by mouth 10 gm of ammonium citrate dissolved in water and flavoured with fruit
juice/lemon.
• Take blood samples after 30, 60, 120 and 180 minutes and determine blood NH3.
• Note: In patients with increased initial levels, give smaller doses, e.g. only 5 grams.
Interpretations
• In normal healthy persons: Little increase is found; blood NH3 levels remaining within normal range.
• In advanced cirrhosis of the liver: Marked rise to twice the initial level or more, exceeding 200 to 300 μg
48 % are seen.
• Considerable increases are also seen when there is a collateral circulation and in patients who have a
portocaval anastomosis.
Determination of glutamine in CS Fluid(An Indirect Liver
Function Test)
PRESENTATION TITLE
• The glutamine is hydrolysed to glutamic acid and NH3 by the action of dilute acid at
100 degree centigrade.
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PRESENTATION TITLE MEGX Test
Principle
Lidocaine is rapidly converted to its primary metabolite monoethyl glycine xylidine
(MEGX) by the hepatic microsomal cytochrome P450 system.
A loss of hepatic cytochrome P450 activity or major changes in hepatic blood flow
(due to portosystemic shunting) result in decreased MEGX formation.
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PRESENTATION TITLE Procedure
• An IV bolus injection of a small lidocaine test dose, 1 mg/ kg, is given.
• Blood sample is taken before the injection.
• Another blood sample is taken 15 or 30 minutes after the injection.
• MEGX is determined in the serum by use of an automated fluorescence
polarisation immuno-assay within about 20 minutes in both the samples.
Interpretations
• In patients with cirrhosis of the liver, the increase of MEGX concentration in serum is
much less marked and decrease value is dependant on disease severity.
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PRESENTATION TITLE Antipyrine breath test
• Antipyrine like lidocaine is also metabolised by cytochrome P450 system.
• When given orally it is absorbed from intestine completely, not bound to plasma
proteins and metabolised by liver only.
Procedure
Interpretations
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PRESENTATION TITLE Alanine aminotransferase (ALT) and Aspartate
transaminase (AST)
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PRESENTATION TITLE Alkaline phosphatase aminotransferase (ALP)
• Normal range for serum ALP as per King-Armströng method is 3 to 13 KA Units/100 ml (23 to 92
IU/L).
• It is increased in both infectious hepatitis (viral hepatitis) and posthepatic jaundice (extrahepatic
obstruction) but the rise is usually much greater in cases of obstructive jaundice.
• Very high values are occasionally found in certain liver diseases, e.g. xanthomatous biliary cirrhosis
in which there is no extrahepatic obstruction.
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PRESENTATION TITLE Serum Lactate Dehydrogenase (LDH)
• Increase in total LDH level is seen in hemolytic anemias, hepatocellular
damage, muscular dystrophy, carcinomas, leukemias, and any condition
which causes necrosis of body cells.
• Thus both NTP & ALP behave similarly in cases of hepatobiliary disease.
MIRJAM NILSSON
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